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Official websites use. Share sensitive information only on official, secure websites. Correspondence: Abdolghaffar Ownagh. There are few studies on Coxiella burnetii Cb as a causative agent of Q fever in dairy products in Iran. The prevalence of Cb was studied by polymerase chain reaction PCR method in Kope pot cheese and cattle milk collected from West Azerbaijan province, Iran. A total number of Kope cheese and milk samples were collected during the year The results showed that There was a significant difference in cheese and milk contaminations with Cb among the defined age groups as well as regional and seasonal variations. It was concluded that Kope cheese and cattle milk are important sources of Cb and should be considered as important risk factors in the epidemiology of Q fever disease in public health. Coxiella burnetii Cb is an intra-cellular Gram-negative pathogenic bacterium, being a causative agent of Q fever. A small cell type with dense chromatin has been considered as a form of extra-cellular survival indicating a remarkable resistance feature to environmental stress factors such as desiccation and heat. The important issues of the risk profile include investigation into the presence of Cb in livestock, milk and dairy products, Cb surviving possibility during the processing of dairy products, pathogenicity of Cb through the inhalation transmission and oral route extremely in accordance with the studies from and , and the way of consumption patterns. Eukaryotic cell is particularly needed to achieve the growth of Cb being discussed as an obligate intra-cellular bacterium. Transmission of infected aerosols probably happens through the human inhalation. Coxiella burnetii is considered to be the most important pathogen frequently detected in ticks. Moreover, their role regarding human infection appears to be ill-defined. Since the epidemic in Netherlands, the global investigation into the field of Q fever attracted considerable interest. Recent findings regarding above issue were in Michigan, United States, consisting five cases. Kope pot cheese is a traditional cheese with the highest production in Greece, Turkey, and Iran named as a Kope or Coupe cheese in the northwest of country. Enhancing the sensitivity of Cb identification has been performed through polymerase chain reaction PCR utilizing particular primers targeting transposon-like element sequence IS of the agent. Sample collection. The study was carried out on samples of Kope cheese collected from cheese makers in the northwest of Iran and cattle milk samples provided from the south of West Azerbaijan province, Iran, in Fig. The animals in the study were grouped based on age into four different groups , , , and over 8 years old. Samples were collected aseptically, placed in a cooler box with ice packs, and immediately transferred to the laboratory. DNA extraction. To extract DNA from milk, 1. The procedure was performed according to the method outlined by De Medici et al. Coxiella burnetii touchdown- nested PCR. To perform the steps in the first stage, the method of Parisi et al. The PCR reactions In the Trans-PCR stage, PCR touchdown was used to optimize, reduce contaminants, remove inhibitors, and also increase the specificity and sensitivity of the reaction. The temperature program of the touchdown method is shown in Table 1. The primers sequences were previously described by Berri et al. Nested-PCR was performed according to the method described by Khademi et al. All conditions of this step such as mixture of PCR reagents, time and temperature schedule were performed according to the Table 1. In this research, bacterial DNA was directly extracted from Kope cheese, since Coxiella cannot grow on usual microbiological media and needs specific cell cultures in biosafety level 3 laboratories. The PCR products for both stages were electrophoresed on a 2. Statistical analysis. The p value less than 0. Coxiella burnetii detection. Out of a total of samples of cattle milk and Kope cheese, samples were positive for the IS gene. Coxiella burnetii prevalence. The prevalence of Cb in all cattle milk and Kope cheese samples was It was found that the prevalence of Cb in cattle milk and Kope cheese samples were Prevalence of Coxiella burnetii in cattle milk collected from dairy farms and contamination of Kope cheese samples collected from West Azerbaijan province, Iran. DNA sequencing. Despite the fact that the infectious and heat resistant pathogen has been reported in cheese from other countries, the content mentioned above was the preliminary discussion regarding the aforesaid pathogen in a ready-to-eat raw-milk Kope cheese in Iran. The Q fever is among the most commonly discussed diseases in animals being attracted by serological and molecular investigations. Therefore, infection originated from consumption of unpasteurized cow milk could be attributed to excreted Cb into the milk. It has not yet established whether consumption of cheese causes the infection or not. Numerous investigations have revealed Cb DNA in milk and derived products, including cheese, cream, butter, and yoghurt from cows, goats, and sheep. Molecular studies have suggested that the Cb genotypes predominating in dairy products are the same as those infect dairy cattle. This study focused on Kope cheese produced in the northwest of Iran, and the milk of cattle living in the south of West Azerbaijan province, Iran; the mentioned region has the potential of cattle breeding. The analysis of this study demonstrated that A serological survey of a cohort of goat farmers and workers, and their contacts, involved in an outbreak of Q fever in the Canadian province of Newfoundland, identified the consumption of a cheese made with pasteurized goat milk as a significant independent risk factor for infection. A study in the United States showed the occurrences of Similar results have also been reported from England, Bulgaria, Slovakia, and Spain. For instance, the rates of contamination in the central Chaharmahal and Bakhtiari province , southern Jahrom , West Azerbaijan, northwest Bonab and southwest Khuzestan province areas of Iran were 6. The most important reasons that could be cited for the difference reported in the prevalence of Cb in dairy products in different parts of the world are the diversities in climate and environments of the geographical areas, the type of survey, the type and number of samples, and the season in which sampling took place. Taken together, the results of the study highlighted that the age is crucially associated with. Cb in the cattle milk. The current study corroborates the reported results showing the considerable risk factor of age in shedding of Cb into the cattle milk. The results of the present study showed that there was a significant regional variation in the shedding of Q fever agent in raw milk. Therefore, it might be speculated that the population of buffalo, cattle and sheep shedding the bacterium will increase the positive samples. A satisfactory explanation for the potential of Cb in forming a spore-like environment is resistant small colony variant type of the bacterium. This factor could be responsible for survival of bacteria in milk and dairy products in a long-term period. In contradiction with unpasteurized milk, the revealed results were in line with Cb scarce viability detected in unpasteurized cheese. Moreover, in contrast with unpasteurized cheese, the findings confirmed the vital epidemiological evidence provided by unpasteurized milk for human cases. Detection of Cb viability in hard cheese after eight months of ripening process would be a worst-case scenario. Viable Cb was proven in raw cheese by culture in Vero cells and inoculation in mice. There has been disagreement concerning application of milk heat treatment on Kope cheese production. Therefore, it is recommended that minimum conditions should be undertaken for farmers manufacturing cheese from unpasteurized milk in the following areas: Positive individuals or herds identification through serological or PCR examinations, keeping the herds in closed form, health certification requirements when buying animals, immunization, prenatal separation and removal of placentas and fetal complications. The Kope cheese is considered as a minimally processed food. Although several authors have called into question the determination of Cb as a foodborne pathogen, this work led us to conclude that Cb risk to humans through the use of raw milk and dairy products could be considered significant. We gratefully acknowledge the sponsorship DVM proposal no. Thanks are also due to Mr. Ali Kazemnia and Dr. Peyman Khademi giving us much valuable technical assistance in the early stages of this work. As a library, NLM provides access to scientific literature. Vet Res Forum. Find articles by Kosar Mokarizadeh. Find articles by Addolghaffar Ownagh. Find articles by Hossein Tajik. Received Dec 12; Accepted May 16; Issue date All rights reserved. Open in a new tab. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel. Touchdown and Trans-PCR.

Assessment of Genetic Diversity and Population Structure in Iranian Cannabis Germplasm

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